ABCA4 p.Thr970Ala
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PMID: 16533065
[PubMed]
Biswas-Fiss EE et al: "Interaction of the nucleotide binding domains and regulation of the ATPase activity of the human retina specific ABC transporter, ABCR."
No.
Sentence
Comment
74
The following complementary oligonucleotides were used as mutagenic primers to generate the mutant NBD1 and NBD2 proteins: NBD1Pro940Arg (5'-GGT AAA GAT TTT TGA GCG CTG TGG CCG GCC AGC TG-3'), NBD1 Walker A, K969A, T970A (5'-CAA TGG AGC TGG GGC AGC CAC CAC CTT GTC CAT CC-3'), NBD2 Walker A, K1978A, T1979A (5'-GAA TGG TGC CGG CGC AGC AAC CAC ATT CAA GAT GC-3'), and NBD2 R2038W (5'-CTT TAC CTT TAT GCC AGG CTT CGA GGT GTA CCA GC-3').
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ABCA4 p.Thr970Ala 16533065:74:215
status: NEW111 Lanes (from left to right): 1, NBD1WT; 2, NBD1P940R; 3, NBD1K969A,T970A; 4, NBD2WT; 5, NBD2R2038W; 6, NBD2K1978A,T1979A.
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ABCA4 p.Thr970Ala 16533065:111:66
status: NEW199 The affinity of interaction decreased under all conditions examined, as determined by comparison of the Kd values (Table Table 2: Modulation of ATPase Activities of NBD1‚NBD2 Complexes by Site-Specific Mutations NBD1‚NBD2 mutation domain NBD1‚NBD2 ATPasea (pmol/min) NBD1‚NBD2 ATPaseexptl (pmol/min) NBD1‚NBD2 ATPasecalcd b (pmol/min) motif/disease NBD1mut‚NBD2wt K969A, T970A NBD1 1.9 ( 0.4/5.3 ( 0.1 5.1 ( 0.1 7.2 synthetic NBD1wt‚NBD2mut K1978A, 1979A NBD2 3.1 ( 0.2/1.6 ( 0.3 1.5 ( 0.2 4.7 synthetic NBD1mut‚NBD2wt P940R NBD1 1.9 ( 0.3/5.2 ( 0.2 4.3 ( 0.1 7.1 AMD NBD1wt‚NBD2mut R2038W NBD2 3.1 ( 0.1/2.0 ( 0.2 4.4 ( 0.2 5.1 STGD NBD1wt‚NBD2wt wild type N/A 3.0 ( 0.3/5.4 ( 0.1 5.30 ( 0.3 8.4 none a Experimental ATPase activity corresponding to 6 pmol of the individual domains, NBD1 and NBD2, respectively.
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ABCA4 p.Thr970Ala 16533065:199:413
status: NEW224 The ATPase assay was carried out as described under Experimental Procedures at 37 °C for 60 min. A theoretical curve is shown, indicating the results expected if the activity of NBD1P940R‚NBD2 were additive (]) of that obtained for the individual domains. Each point represents the average of three independent experiments. FIGURE 6: Effect of NBD1 Walker A mutations on modulation of ATPase of the NBD1‚NBD2 protein complex. Protein titration of the ATPase activity of purified NBD2WT (∆), NBD1K969A,T970A (0), NBD1WT (9), and the NBD1K969A,T970A‚NBD2 complex (O).
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ABCA4 p.Thr970Ala 16533065:224:527
status: NEW